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BACKGROUND: Environmental stress can induce oxidative stress in Apis cerana cerana, leading to cellular oxidative damage, reduced vitality, and even death. Currently, owing to an incomplete understanding of the molecular mechanisms by which A. cerana cerana resists oxidative damage, there is no available method to mitigate the risk of this type of damage. Cyclin plays an important role in cell stress resistance. The aim of this study was to explore the in vivo protection of cyclin H against oxidative damage induced by abiotic stress in A. cerana cerana and clarify the mechanism of action. We isolated and identified the AccCyclin H gene in A. cerana cerana and analysed its responses to different exogenous stresses. RESULTS: The results showed that different oxidative stressors can induce or inhibit the expression of AccCyclin H. After RNA‐interference‐mediated AccCyclin H silencing, the activity of antioxidant‐related genes and related enzymes was inhibited, and trehalose metabolism was reduced. AccCyclin H gene silencing reduced A. cerana cerana high‐temperature tolerance. Exogenous trehalose supplementation enhanced the total antioxidant capacity of A. cerana cerana, reduced the accumulation of oxidants, and improved the viability of A. cerana cerana under high‐temperature stress. CONCLUSION: Our findings suggest that trehalose can alleviate adverse stress and that AccCyclin H may participate in oxidative stress reactions by regulating trehalose metabolism. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The intestinal microbial community composition of different bee species typically has host specificity, yet little is known about the underlying formation mechanism. There are signs that dietary habits vary in different bee species, suggesting that there may be close relationships between dietary habits and intestinal microorganisms. We explored this hypothesis by comparing the dietary habits and gut microbiota of two common bee species (Apis mellifera L. and Apis cerana cerana) in China. Bee bread and midgut samples from wild and laboratory-reared bees were collected, and the differences in intestinal microbial community composition and growth and development before and after the change in dietary habits of different bee species were compared. We found that the two sympatric species had different dietary specializations and similar metagenomic diversities. The microbiota composition differed between the two species. Moreover, we revealed that changes in native dietary habits destroyed the intestinal microbiota community composition, negatively affecting the growth and development of honeybees. [ABSTRACT FROM AUTHOR]

Odorant-binding proteins (OBPs) play an important role in odour perception and transport in insects. However, little is known about whether OBPs perform other functions in insects, particularly in Apis cerana cerana. Within this study, an OBP gene (AccOBP10) was isolated and identified from A. c. cerana. Both homology and phylogenetic relationship analyses indicated that the amino acid sequence of AccOBP10 had a high degree of sequence identity with other members of the gene family. Analysis of quantitative real-time PCR (qRT-PCR) showed that AccOBP10 mRNA was expressed at higher levels in the venom gland than in other tissues. The mRNA transcript expression of AccOBP10 was upregulated by low temperature (4°C), hydrogen peroxide (H2O2), pyridaben, methomyl and imidacloprid but downregulated by heat (42°C), ultraviolet light, vitamin C, mercuric chloride, cadmium chloride, paraquat and phoxim. Expression of AccOBP10 under abiotic stress was analysed by western blotting, and the results were consistent with those of qRT-PCR. And as a further study of AccOBP10 function, we demonstrated that knockdown of AccOBP10 by RNA interference could slightly increase the expression levels of some stress-related genes. Collectively, these results suggest that AccOBP10 is mainly involved in the response to stress conditions. [ABSTRACT FROM AUTHOR]

Cytochrome P450 monooxygenases (P450s) play significant roles in protecting organisms from abiotic stress damage. Here, we report the sequence and characterization of a P450s gene (AccCYP4AV1), isolated from Apis cerana cerana Fabricius. The open reading frame of AccCYP4AV1 is 1506 base pairs long and encodes a predicted protein of 501 amino acids and 57.84 kDa, with an isoelectric point of 8.67. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis indicated that AccCYP4AV1 is more highly expressed in the midgut than in other tissues. In addition, the highest expression occurs in newly emerged adult workers, followed by the first instar of the larval stage. In addition, the expression of the AccCYP4AV1 was upregulated by low temperature (4 °C), ultraviolet radiation, hydrogen peroxide, paraquat, and dichlorvos treatments. In contrast, AccCYP4AV1 transcription was downregulated by other abiotic stress conditions: exposure to increased temperature (44 °C), deltamethrin, cadmium chloride, and mercury (II) chloride. Moreover, when AccCYP4AV1 was knocked-down by RNA interference, the results suggested that multiple antioxidant genes (AccsHSP22.6, AccSOD2, AccTpx1, and AccTpx4) were downregulated and antioxidant genes AccGSTO1 and AccTrx1 were upregulated. The activity levels of peroxidase and catalase were upregulated in the AccCYP4AV1-knocked-down samples, compared with those in the control groups. These findings suggest that the AccCYP4AV1 protein might be involved in the defense against abiotic stress damage. [ABSTRACT FROM AUTHOR]

UDP-glucuronosyltransferases (UGTs), being multifunctional detoxification enzymes, play a major role in the process of resistance to various pesticides in insects. However, the mechanism underlying the molecular regulation of pesticide resistance remains unclear, especially in Apis cerana cerana. In this study, all of the UGTs in Apis cerana cerana (AccUGT) have been identified through the multiple alignment and phylogenetic analysis. Expression of AccUGT genes under different pesticides, and antioxidant genes after silencing of AccUGT2B20-like , were detected by qRT-PCR. The resistance of overexpressed AccUGT2B20-like to oxidative stress was investigated by an Escherichia coli overexpression system. Also, antioxidant-related enzyme activity was detected after silencing of the AccUGT2B20-like gene. Expression pattern analysis showed that almost all UGT genes were upregulated under different pesticide treatments. This result indicated that AccUGTs participate in the detoxification process of pesticides. AccUGT2B20-like was the major gene because it was more highly induced than the others. Overexpression of AccUGT2B20-like in E. coli could effectively improve oxidative stress resistance. Specifically, silencing the AccUGT2B20-like gene increased oxidative stress by repressing the expression of oxidation-related genes, decreasing antioxidant-related enzyme activity, and increasing malondialdehyde concentration. Taken together, our results indicate that AccUGTs are involved in pesticide resistance, among which, AccUGT2B20-like contributes to the detoxification of pesticides by eliminating oxidative stress in Apis cerana cerana. This study explains the molecular basis for the resistance of bees to pesticides and provides an important safeguard for maintaining ecological balance. [ABSTRACT FROM AUTHOR]

Cytochrome P450 monooxygenases (P450s) are widely distributed multifunctional enzymes that play crucial roles in insecticide detoxification or activation. In this study, to ascertain the molecular mechanisms of P450s in the detoxification of Chinese honeybees, Apis cerana cerana Fabricius (A. c. cerana), we isolated and characterized four new P450 genes (Acc301A1 , Acc303A1 , Acc306A1, and Acc315A1). The open reading frames of the four genes are 1263 to 1608 bp in length and encode four predicted polypeptides of 499 to 517 amino acids in length. Real-time quantitative PCR (RT-qPCR) results showed that expression of all four genes was observed in all developmental stages. In addition, Western blot assays further indicated the RT-qPCR results that showed that the four genes were induced by pesticide (thiamethoxam, deltamethrin, dichlorovos, and paraquat) treatments. Furthermore, we also used double-stranded RNA-mediated RNA interference to investigate the functions of Acc301A1 , Acc303A1 , and Acc306A1 in the antioxidant defense of honeybees. RNA interference targeting Acc301A1 , Acc303A1 , and Acc306A1 significantly increased the mortality rate of A. c. cerana upon pesticide treatment. These results provide important evidence about the role of the four P450 genes involved in detoxification. [ABSTRACT FROM AUTHOR]

The effects of insecticides on bee health are a topic of intensive research. Although abamectin is toxic to bees, the molecular impact of abamectin needs to be clarified. Here, we found that Apis cerana cerana exhibited a higher mortality rate when exposed to abamectin than Apis mellifera ligustica. In addition, A. cerana cerana had markedly higher numbers of differentially expressed genes (DEGs), differentially expressed proteins (DEPs) and differentially expressed metabolites (DEMs) than A. mellifera ligustica during exposure to abamectin. These results indicate that abamectin exposure exerts stronger effects on A. cerana cerana than on A. mellifera ligustica. In addition, six DEGs, two DEPs and two DEMs overlapped between the two bee species under abamectin exposure; however, some genes or proteins from the zinc finger protein, superoxide dismutase and peroxiredoxin families and the energy metabolism pathway were only unregulated in A. cerana cerana , which indicates a significant difference in the impact of abamectin on the two bee species. Despite these differences, several of the same gene families, such as heat shock proteins, cytochrome P450, odorant-binding proteins and cuticle proteins, and pathways, including the carbohydrate metabolism, immune system, lipid metabolism, amino acid metabolism, sensory system, locomotion and development pathways, were influenced by abamectin exposure in both A. cerana cerana and A. mellifera ligustica. Together, our results indicate that abamectin causes adverse effects on bees and thus poses a risk to bee populations and that abamectin exposure affects A. cerana cerana more strongly than A. mellifera ligustica. These findings improve our understanding of the behavioural and physiological effects of abamectin on bees. • Abamectin disturbs the transcription, translation and metabolism networks of bees. • Abamectin affects A. cerana cerana more strongly than A. mellifera ligustica. • Many genes, proteins and metabolites play key roles in response to abamectin in bees. [ABSTRACT FROM AUTHOR]

Environmental pollution has gained negative attention in recent years. The pesticides and heavy metals are top list of environmental toxicants directly endangering the survival and development of Apis cerana cerana. Cyclin-dependent kinases (CDKs) are heteromeric serine/threonine kinases that participate in cell cycle regulation and have a vital role in pesticide and heavy metal stress in Apis cerana cerana. In this experiment, we filtered out CDK8 gene from Apis cerana cerana (AccCDK8) and investigated its functions of pesticide and heavy metals resistance. Sequence analysis indicated that AccCDK8 is highly homologous to multiple CDK8s and contains a highly conserved CDK active site sequence. Phylogenetic analysis showed that AmCDK8 and AccCDK8 were closely related evolutionarily in Apis mellifera. Transcriptome analysis revealed that AccCDK8 expression was differentially affected after exposure to pesticide and heavy metal stresses. This indicates that AccCDK8 has a significant role in the resistance of Apis cerana cerana to pesticide and heavy metal stresses. It has implications for studying the function of CDK in other insects in response to stress. [Display omitted] • Pesticides and heavy metals affect the survival of Apis cerana cerana. • CDK8 takes part in the oxidative stress response of Apis cerana cerana. • CDK8 is involved in the growth and reproduction of Apis cerana cerana. • AccCDK8 plays an important role in pesticide and heavy metals stresses. • We highlight the effects of some pesticides on the health of Apis cerana cerana. [ABSTRACT FROM AUTHOR]

• Orientation flight activities are related to the development trend of bee colonies. • When the YBR is low and the fluctuation is stable, the bee colony tends to have a better development trend. • Directional flight activities also be short-term colony development. • The changes of bee colonies under different temperatures and periods confirm the regularity of YBR. • When YBR is 0.2, it may be the key point of the change in the development trend of the bee colony. Pollinations as a result of bees collecting pollen from flowers are vital for maintaining the subtle balance of the earth's ecological system. However, apiculture is facing a crisis owing to collapsing honey bee colonies, yet the reasons are still unclear. As young bees are the future backbone of a colony, their behaviors could reveal the development of the colony and the problems causing colony collapse. In this study, we used Internet of Things technology to monitor the data of eight colonies of Apis cerana continuously over three years. We obtained more than 10,000 h of monitoring data to analyze the relationship between the orientation flight activities of young bees and changes in honeycomb weight, time, and local ambient temperature. The results indicated that the young bee ratio was closely associated with the trend in colony development. When the ratio of young bees participating in the orientation flight was low and stable, the colony size was likely to be maintained; in contrast, when the ratio was high and fluctuating considerably, the bee colony tended to decline. This finding can be used to predict the development trend of bee colonies and explore further quantitative factors that trigger abnormal colony losses. [ABSTRACT FROM AUTHOR]

Previous studies examining the functions of cyclin-dependent kinases (CDKs) have mainly focused on the regulation of the cell cycle. Recent studies have found that cyclin-dependent kinase 7 (CDK7) and cyclin-dependent kinase 9 (CDK9) play important roles in cell stress, metabolism of toxic substances and maintaining the stability of the internal environment. Here, we found that under stress conditions, the transcription and protein expression of AccCDK7 and AccCDK9 were induced to varying degrees. Meanwhile, the silencing of AccCDK7 and AccCDK9 also affected the expression of antioxidant genes and the activity of antioxidant enzymes, and reduced the survival rate of bees under high temperature stress. Furthermore, the exogenous overexpression of AccCDK7 and AccCDK9 improved the viability of yeast under stress conditions. Therefore, AccCDK7 and AccCDK9 may play roles in A.cerana cerana resistance to oxidative stress caused by external stimuli, potentially revealing a new mechanism of the honeybee response to oxidative stress. • Isolation of AccCDK7 and AccCDK9 from Apis cerana cerana. • The expression of AccCDK7 and AccCDK9 is induced by oxidative stress. • AccCDK7 and AccCDK9 can enhance the stress resistance of E. coli and yeast. • RNA interference experiments suggested that AccCDK7 and AccCDK9 is important for resistance against stresses. [ABSTRACT FROM AUTHOR]

In eukaryotes, cytochrome c oxidase (COX) is a multimeric protein complex that is the last enzyme in the respiratory electron transport chain of mitochondria. Syntheses of cytochrome c oxidase (SCO) proteins are copper-donor chaperones involved in metalation of the CuA redox center of COX. However, its other precise actions are not yet understood. Here, we report the characterization of AccSCO2 from Apis cerana cerana ( Acc ). Our data showed that AccSCO2 expression was induced by cold (4 °C), CdCl 2 , HgCl 2 , ultraviolet (UV) light, and H 2 O 2 and was inhibited by different pesticide treatments. In addition, a disc diffusion assay of recombinant AccSCO2, AccSCO2-R1, and AccSCO2-R2 proteins showed that they played a functional role in protecting cells from oxidative stress involved in copper-dependent manner. Further, following knockdown of AccSCO2 in A. cerana cerana using RNA interference (RNAi), the expression levels of most antioxidant genes ( AccGSTD , AccGSTO1 , AccGSTS4 , AccSOD1 , AccSOD2 , etc.) were significantly decreased in the AccSCO2 -silenced bees compared with the control bees. Moreover, the antioxidant enzymatic activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) were all lower in the silenced bees than in the control bees. Finally, the in vivo activity of COX was measured after AccSCO2 knockdown, which revealed a strong reduction in COX activity in the silenced bees. Thus, we hypothesize that AccSCO2 plays important roles in cellular stress responses and anti-oxidative processes, which help to regulate the production of mitochondrial reactive oxygen species and/or the impairment of mitochondrial activity under oxidative stress. [ABSTRACT FROM AUTHOR]

Insect cytochrome P450 monooxygenases (P450s or CYPs) perform important functions in the metabolic detoxification of both endogenous and exogenous substrates. However, the mechanism of action of the P450 genes in bees is unclear. In this study, we investigated the effects of AccCYP6k1 on the metabolism and detoxification of Apis cerana cerana. Spatiotemporal expression profiling revealed that the expression of AccCYP6k1 was the highest in foragers (A15) and was mainly expressed in the leg, midgut and head. RT–qPCR results showed that AccCYP6k1 exhibited different expression patterns following exposure to xenobiotics. In addition, silencing AccCYP6k1 increased the pesticides sensitivity and affected the detoxification system and antioxidant process of A. cerana cerana. In brief, the induced expression of AccCYP6k1 is related to the resistance of A. cerana cerana , while knockdown AccCYP6k1 affect the pesticides resistance and metabolic detoxification system of A. cerana cerana. These findings not only support the theoretical basis of metabolic detoxification in bees but also provide a better understanding of P450-mediated resistance to pesticides in insects. [Display omitted] • The level of inducible expression of AccCYP6k1 is related to pesticide resistance. • Silencing AccCYP6k1 increased the sensitivity of Apis cerana cerana to abamectin and imidacloprid. • Silencing AccCYP6k1 had adverse effects on the detoxification system of Apis cerana cerana. [ABSTRACT FROM AUTHOR]

Tyrosine aminotransferase (TATN) is the first enzyme involved in the metabolic degradation of tyrosine, and it plays an important role in tyrosine detoxification and helps the body resist oxidative damage. However, the function of TATN in Apis cerana cerana (A. c. cerana) remains unclear. To explore the role of TATN in the response to pesticide and heavy metal stress in A. c. cerana , AccTATN was isolated and identified. AccTATN was highly expressed in the integument and the adult stage. Exposure to multiple pesticides and heavy metal stress upregulated AccTATN expression. RNA interference experiments showed that silencing AccTATN reduced the resistance of A. c. cerana to glyphosate and avermectins stress. The expression of antioxidant-related genes and the activity of antioxidant enzymes were reduced after AccTATN was silenced, leading to the accumulation of oxidative damage. Overexpression of the recombinant AccTATN protein in a prokaryotic system also confirmed its role in heavy metal stress and improved antioxidant capacity. Our study showed that AccTATN may promote resistance to pesticide and heavy metal stress by regulating the antioxidant capacity of A. c. cerana. This study provides a valuable theoretical basis for A. c. cerana conservation. [Display omitted] • AccTATN gene was isolated and characterized from (A. c. cerana) • Pesticide and heavy metal stress can affect the expression of AccTATN. • AccTATN overexpression enhanced the tolerance of E. coli cells to oxidative stress. • AccTATN knockdown disrupted the original redox balance and caused oxidative stress. • AccTATN promotes A. c. cerana to resist pesticide and heavy metal stress. [ABSTRACT FROM AUTHOR]

Apis cerana cerana is a native bee species in China and plays a key role in agricultural production and ecological balance. However, the growth and development of Apis cerana cerana has not been smooth, and pesticide and heavy metal stress are key factors that have forced a dramatic decline in population size. This study was performed with the objective of investigating the role of AccCDK20 and AccCDKN1 in honey bee resistance to pesticide and heavy metal stress. RT–qPCR analysis revealed that AccCDK20 transcript levels were highest in brown-eyed pupae and AccCDKN1 transcript levels were highest in 1-day-old worker bees. In different tissues and body parts of adult bees, AccCDK20 transcript levels were highest in the head, and AccCDKN1 transcript levels were highest in the thorax. It was further observed that environmental stress can affect the transcript levels of the AccCDK20 and AccCDKN1 genes. Silencing of the AccCDK20 and AccCDKN1 genes resulted in altered activities of antioxidant-related genes and antioxidant-related enzymes. AccCDK20 and AccCDKN1 transcript levels were upregulated under glyphosate stress, and silencing of the genes resulted in reduced resistance to glyphosate and greatly increased mortality in Apis cerana cerana. In addition, gene function was verified by in vitro repression assays. Overexpression of the AccCDK20 and AccCDKN1 proteins in E. coli cells increased the resistance to ROS damage induced by CHP. In conclusion, AccCDK20 and AccCDKN1 play an indispensable role in honey bee resistance to pesticide and heavy metal stress. [Display omitted] • Silencing AccCDK20 and AccCDKN1 affects the oxidative system. • AccCDK20 and AccCDKN1 promote honey bee resistance to glyphosate stress. • Overexpression of AccCDK20 and AccCDKN1 enhances the antioxidant capacity of E. coli. [ABSTRACT FROM AUTHOR]

The cyclin-dependent kinase (CDK) protein family plays an important role in regulating life functions, such as the cell cycle and metabolism. This study reports the first cloning and functional analysis of A. cerana cerana CDK1 (AccCDK1). The distribution profile of AccCDK1 in different developmental periods and different tissues was determined. The experimental results showed that the distribution of AccCDK1 was tissue-specific. AccCDK1 distribution at the transcriptional and translational levels was affected by stress conditions induced by H 2 O 2 , UV, HgCl 2 , CdCl 2 , extreme temperatures (4 °C, 44 °C) and pesticides (avermectin, lambda-cyhalothrin, haloxyfop- R -methyl, and glyphosate), which resulted in changes in the expression levels. These results suggest that AccCDK1 may have an important part to play in honey bee resistance to stress. The expression of a recombinant AccCDK1 protein in vitro enhanced the antistress capacities of E. coli and yeast, which suggests that AccCDK1 is related to the stress response. When AccCDK1 was silenced, the expression of some antioxidant genes was downregulated, and the enzymatic potencies of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) were reduced, which suggests that AccCDK1 takes part in the body's resistance to oxidative stress upon external stimulation by influencing relevant antioxidants. Notably, the survival rate of A. cerana cerana under high-temperature-induced stress decreased after AccCDK1 silencing, which verifies our results. In conclusion, we found that AccCDK1 played an indispensable function in resisting oxidative stress and maintaining normal cellular functions. [Display omitted] • Oxidative stress can affect the expression of cyclin-dependent kinase1. • AccCDK1 silencing affects cellular antioxidant genes and enzymes. • AccCDK1 promotes the survival of eukaryotic and prokaryotic cells under stress. • AccCDK1 promotes bees to resist high temperature stress. [ABSTRACT FROM AUTHOR]